Diagnosis and treatment of cancer using anti-desmoglein-3 antibody

An antibody and protein technology, applied in the direction of antibodies, anti-tumor drugs, cells modified by introducing foreign genetic material, etc., can solve the problems of low diagnostic positive rate and low disease cure rate

Inactive Publication Date: 2009-08-12
FORERUNNER PHARMA RES CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the positive rate of diagnosis and the cure rate of the disease are still low. In order to apply molecularly targeted therapy using antibodies exhibiting the above-mentioned activities to the treatment of lung cancer that still has a chance of complete remission, it is strongly desired to express tumor-specific expression in lung cancer cells. Molecules are identified and targeted to produce antibodies that can exert the desired activity

Method used

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  • Diagnosis and treatment of cancer using anti-desmoglein-3 antibody
  • Diagnosis and treatment of cancer using anti-desmoglein-3 antibody
  • Diagnosis and treatment of cancer using anti-desmoglein-3 antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0473] [Example 1] DSG 3 mRNA expression analysis in various cancers

[0474] For DSG3 gene expression analysis, microarrays were used. In order to investigate genes highly expressed in cancer cells, total RNA prepared from various RNAs shown in Tables 1 and 2 and various excised tissues by ISOGEN (manufactured by Nippon Gene) according to a conventional method was used. More specifically, 10 μg of total RNA was used to supply gene chip U-133A (manufactured by Affymetrix), and gene expression analysis was carried out according to the expression analysis technical manual (manufactured by Affymetrix). In the analysis of lung adenocarcinoma and hepatocellular carcinoma, a total of 10 μg of total RNA was analyzed from 12 cases of lung adenocarcinoma and 3 cases of hepatocellular carcinoma (Table 1).

[0475] [Table 1]

[0476] organize source whole brain Clontech 64020-1 lung 1 case of clinical sample trachea Clontech 64091-1 heart Ambion ...

Embodiment 2

[0484] [Example 2] Immunohistological staining of DSG3 in lung squamous cell carcinoma

[0485] Transcription of the DSG3 gene is increased in cancer cells, particularly lung squamous cell carcinoma cells. Accordingly, in order to confirm the expression of the DSG3 protein, immunohistostaining analysis was performed.

[0486] Each test sample was prepared as a fixed paraffin-embedded specimen, cut into a 4 μm thin section, and the section was stuck on a glass slide, and then left at 37° C. for about 16 hours to make it fully dry. The section was deparaffinized by immersing in 100% xylene for 5 minutes three times, soaking in 100% ethanol for 5 minutes three times, and further immersing in 70% ethanol for 5 minutes to perform hydrophilic treatment. After washing with 50 mM TBS buffer for 5 minutes, repeated three times, the slice was treated in citrate buffer (10 mM, pH 7.0) at 120° C. for 10 minutes to activate the antigen in the slice. The section after the antigen activatio...

Embodiment 3

[0488] [Example 3] Preparation of anti-DSG3 antibody

[0489] 3-1) Cloning of full-length cDNA encoding human DSG3

[0490] The full-length cDNA encoding human DSG3 can be obtained by PCR amplification using Human Small Intestine Marathon-Ready cDNA (CLONTECH) as a template. That is, 50 μL containing 2 μL cDNA, 1 μL sense primer (SEQ ID NO.37), 1 μL antisense primer (SEQ ID NO.38), 5 μL 10×KOD-Plus buffer, 5 μL 2mMdNTPs, 2 μL 25mM MgSO 4 , 1 μL of KOD-Plus reaction solution, and carried out the PCR reaction continuously under the following conditions: the reaction cycle consisting of 94°C for 15 seconds and 70°C for 2 minutes was carried out 5 times, and the reaction cycle of 94°C for 15 seconds and 68°C for 2 minutes was performed 5 times. The reaction cycle consisting of the reaction of 94° C. for 15 seconds and the reaction of 66° C. for 2 minutes was carried out 28 times. The amplification product obtained by the above PCR reaction was inserted into pGEM-T easy using p...

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Abstract

Disclosed is a method for the diagnosis of cancer, which is characterized by detecting a DSG3 protein. It is found that the increase in the expression level of DSG3 is observed in lung cancer highly frequently at a gene or protein level. The method can be effected by using an antibody capable of recognizing a DSG3 protein. Also disclosed are a pharmaceutical composition, a cell proliferation inhibitor and an anti-cancer agent, each comprising an antibody capable of binding to DSG3 as an active ingredient. Further disclosed a method for inducing the cytopathy in a DSG3-expressing cell and a method for inhibiting the proliferation of a DSG3-expressing cell, each by contacting the DSG3-expressing cell with an antibody capable of binding to DSG3.

Description

technical field [0001] The present invention relates to methods for diagnosis and treatment of cancer, as well as cell proliferation inhibitors and anticancer drugs. Background technique [0002] Demoglein 3 (Demoglein3) (hereinafter referred to as DSG3 in this specification) molecule is caused by suffering from the autoimmune blister formation disease of skin and mucous membrane—pemphigus vulgaris (Pemphigus vulgaris, hereinafter referred to as PV in this specification). ) in the serum of patients with autoantibodies, and using the autoantibodies to perform immunoprecipitation on keratinocyte extracts, a glycoprotein with a molecular weight of 130 kDa was identified for the first time and named as PV antigen (hereinafter referred to as PVA in this specification) (non-patented Document 1 and J. Clin. Invest. 74, 313-320, 1984). Then, antibody molecules reacting with the above-mentioned 130 kDa protein were isolated from PV patient serum by affinity purification. Next, usin...

Claims

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Application Information

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IPC IPC(8): A61K39/395A61P1/04A61P1/18A61P11/00A61P35/00A61P43/00C12N15/09G01N33/574C12N5/10
Inventor 油谷浩幸石川俊平伊藤浩孝中野清孝川合重人
Owner FORERUNNER PHARMA RES CO LTD
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